Anthocyanins in cowpea [Vigna unguiculata (L.) Walp. ssp. unguiculata]

The aim of this study was to isolate and identify the anthocyanins in the black seed coated cowpea [Vigna unguiculata (L.) Walp. ssp. unguiculata] using reverse phase C-18 open column chromatography and high performance liquid chromatography (HPLC) with diode array detection and electro spray ionization/mass spectrometry (DAD-ESI/MS) analysis, respectively. Anthocyanins were extracted from the coat of black cowpea with 1% trifluoroacetic acid (TFA) in methanol, isolated by RP-C-18 column chromatography, and their structures elucidated by 1D and 2D nuclear magnetic resonance (NMR) spectroscopy. The isolated anthocyanins were characterized as delphinidin-3-O-glucoside (2) and cyanidin-3-O-glucoside (4). Furthermore, 5 minor anthocyanins were detected and identified as delphinidin-3-O-galactoside (1), cyanidin-3-O-galactoside (3), petunidin-3-O-glucoside (5), peonidin-3-O-glucoside (7), and malvidin-3-O-glucoside (8) based on the fragmentation patterns of HPLC-DADESI/MS analysis.     

Anthocyanin fingerprinting of true bokbunja (Rubus coreanus Miq.) fruit

The interest in black raspberry products has been increasing due to its flavor and potential health benefits. While black raspberries grown in North America are Rubus occidentalis, there has been some confusion regarding the identity of black raspberry grown in Korea (known as bokbunja; Rubus coreanus). As such, there is a need to define the anthocyanin profile of R. coreanus fruit collected from a verified source. We analyzed three genotypes of bokbunja fruit for anthocyanin profiles. While each varied in its anthocyanin proportions and total concentration, bokbunja fruit contained three anthocyanins: cyanidin-3-glucoside, cyanidin-3-rutinoside, and pelargonidin-3-glucoside, and lacked the xylose containing glycosides characteristic of R. occidentalis: cyanidin-3-sambubioside, and cyanidin-3-xylosylrutinoside. Due to mix-ups of bokbunja identity, research claiming to be conducted on bokbunja fruit requires confirmation that the fruit was sourced from a correctly identified plant. The distinct anthocyanin profiles between the two species can be used to confirm plant identity.     

Analysis of Phenolic Compounds of Dabai (Canarium odontophyllum Miq.) Fruits by High-Performance Liquid Chromatography

Dabai (Canarium odontophyllum Miq.) fruit is a popular seasonal indigenous fruit of Sarawak, Malaysia that is always appreciated as an exotic fruit by the local community. The present study was designed to identify and quantify the phenolic compounds of dabai fruits from different divisions of Sarawak. Difference between the purple and red varieties of the fruits was also investigated. Analysis of phenolic compounds was performed by reversed phase high-performance liquid chromatography coupled with diode array detector. Three detection wavelengths (280, 254 and 329 nm) were used to monitor all phenolic compounds (except for anthocyanidins) since they display absorbance maxima at different wavelengths. Characteristic spectra of eluted peaks provide useful information in confirming the identities of peaks. Hydrolysis of phenolic compounds of dabai fruits was insufficient under the milder conditions (30 °C, 2 h) employed thus resulted in unidentified peaks. Two phenolic acids (ellagic and vanillic acids), five flavonoids (catechin, epicatechin, epicatechin gallate, epigallocatechin gallate, apigenin) and ethyl gallate were detected in the fruits. Furthermore, three anthocyanidins (cyanidin, pelargonidin and delphinidin) and four anthocyanins (malvidin-3,5-di-O-glucoside, cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside and peonidin-3-O-glucoside) were detected. All these phenolic compounds were reported in dabai fruits for the first time. The fruit variety affected the anthocyanidins and anthocyanins profile (p < 0.01) but had little or no effect on the phenolic acids and flavonoids profile of the fruits. Within the same variety, diversity in environmental conditions had an impact on the quantitative differences of phenolic compounds (p < 0.05).     

Positive effects of glycosylated anthocyanin isolated from an edible berry fruit (Acanthopanax sessiliflorum) on its antioxidant activity and color stability

A major chromatographic peak of anthocyanin from ogaja (Acanthopanax sessiliflorum) fruit represented 95.4% of total absorbable compounds at 518 nm. The mass-to-charge ratio (m/z) of this colorant was determined to be 581 by LC-MS, which was identical to the molecular mass of cyanidin-3-O-sambubioside. After acid hydrolysis of the purified anthocyanin fraction, it was structurally confirmed by thin layer chromatography (TLC) and high-performance anion exchange chromatography (HPAEC) analyses. As a result, the sugar moieties linked to cyanidin were glucose and xylose. The purified cyanidin-3-O-sambubioside and cyanidin had greater antioxidant activities than l-ascorbic acid by both DPPH and ABTS methods. Cyanidin-3-O-sambubioside was more stable than cyanidin in the response to heat (80 °C), and both fluorescent and UV light. l-Ascorbic acid had an adverse effect on the color stability of anthocyanin. In this study, we demonstrated that glycosylation on anthocyanidin would provide an effective protection barrier for its color stability against the exposure to various food processing conditions.     

Influence of Dekkera bruxellensis on the contents of anthocyanins,organic acids and volatile phenols of Dão red wine

The anthocyanin, organic acid and volatile phenol compositions of red wine obtained from Touriga Nacional grapes growing in the Dão region (Portugal) were determined by HPLC/DAD, HPLC/UV and GC/FID, respectively. By these means, nine anthocyanic compounds (malvidin-3,5-O-diglucoside, cyanidin-3-O-galactoside, cyanidin-3-O-glucoside, peonidin-3-O-glucoside, malvidin-3-O-glucoside, delphinidin, cyanidin, pelargonidin and malvidin), six organic acids (ketoglutaric, tartaric, malic, quinic, lactic and shikimic acids) and two volatile phenols (4-ethylguaiacol and 4-ethylphenol) were identified and quantified. Malvidin-3-O-glucoside, the pair lactic plus shikimic acids and 4-ethylguaiacol were the main anthocyanin, organic acids and volatile phenol, respectively. The effects of nine different Dekkera bruxellensis strains on these chemical parameters were also evaluated. The results obtained indicate that some strains of D. bruxellensis yeast are able to cause deterioration of red wine from the Dão region during its maturation by the production of volatile phenols, namely 4-ethylphenol.     

Characterisation of phenolic phytochemicals and quality changes related to the harvest times from the leaves of Korean purple perilla (Perilla frutescens)

The objective of this study was to investigate the profiles of phenolic phytochemicals in the leaves of Korean purple perilla (cv. Bora, Perilla fructescens) using reversed-phase C₁₈ column chromatography and HPLC with DAD-ESI/MS analysis. Changes in their contents were also the first reported through eight different harvest times during two months. They were characterised as five anthocyanins and three phenolic acids including cyanidin-3,5-di-O-glucoside (1), cyanidin-3-O-glucoside (2), cyanidin-3-O-(6-O-caffeoyl)glucoside-5-O-glucoside (3), cyanidin-3-O-(6-O-coumaroyl)glucoside-5-O-glucoside (4), cyanidin-3-O-(6-O-coumaroyl)glucoside (5), caffeic acid (6), rosmarinic acid (7), and rosmarinic acid methylester (8). Significant differences were observed between individual and total phytochemical contents, especially, cyanidin-3-O-(6-O-coumaroyl)glucoside-5-O-glucoside (4) and rosmarinic acid (7) exhibited the predominant constituents. Among different harvest times, the highest content was found with 82.473 mg/g on 21st September, while the lowest was 39.000 mg/g on 17th August. These results may be useful in determining the optimal harvest time at which phenolic phytochemicals reaches a maximum level in mid-September.     

Characterisation of anthocyanins in the black soybean (Glycine max L.) by HPLC-DAD-ESI/MS analysis

The aim of this study was to isolate and identify the anthocyanins in the black seed coated soybean (cv. Cheongja 3, Glycine max (L.) Merr.) using reverse phase C-18 open column chromatography and high-performance liquid chromatography (HPLC) with diode array detection and electro spray ionization/mass spectrometry (DAD-ESI/MS) analysis, respectively. Anthocyanins were extracted from the coat of black soybeans with 1% TFA in methanol, isolated by RP-C-18 column chromatography, and their structures elucidated by 1D and 2D NMR spectroscopy. The isolated anthocyanins were characterised as delphinidin-3-O-glucoside (3), cyanidin-3-O-glucoside (5), petunidin-3-O-glucoside (6), pelargonidin-3-O-glucoside (7) and cyanidin (9). Furthermore, four minor anthocyanins were detected and identified as catechin-cyanidin-3-O-glucoside (1), delphinidin-3-O-galactoside (2), cyanidin-3-O-galactoside (4), and peonidin-3-O-glucoside (8) based on the fragmentation patterns of HPLC-DAD-ESI/MS analysis.     

Identification and quantification of anthocyanins from the grains of black rice (Oryza sativa L.) varieties

The purpose of this research was conducted to determine anthocyanin content from the grains of 10 Korean black rice varieties. Moreover, the primary constituents including protein and oil were evaluated. Anthocyanins, cyanidin-3-O-glucoside (1) and peonidin-3-O-glucoside (2), were isolated and elucidated using reversephase C₁₈ chromatography, nuclear magnetic resonance (NMR) spectroscopy, and high performance liquid chromatography (HPLC) with diode array detection and electrospray ionization/mass spectrometry (DAD-ESI/MS). Anthocyanin showed significant differences and cyanidin-3-O-glucoside (1) (52.1±6.3-1,601.0±8.5 μg/g) exhibited a markedly higher content than peonidin-3-O-glucoside (2) (ND-82.6±1.2 μg/g). Among varieties, ‘Heugjinjubyeo’ showed the highest anthocyanin (1: 1,601.0±8.5, 2: 82.6±1.2 μg/g), whereas ‘Heughyang’ was the lowest (1: 52.1±6.3 μg/g, 2: ND). Protein and oil exhibited the minor differences and their contents ranged from 10.7±0.0 to 14.1±0.1% and from 2.1±0.0 to 2.9±0.0%. Overall results suggest that anthocyanin can be a key factor in functional property of black rice and ‘Heugjinjubyeo’ may be very important source concerning nutritional value.     

Optimization of microwave-assisted extraction of anthocyanins in red raspberries and identification of anthocyanin of extracts using high-performance liquid chromatography – mass spectrometry

Anthocyanins (Acys) are naturally occurring compounds that impart color to fruit, vegetables, and plants. The extraction of Acys from red raspberry (Rubus idaeus L. var. Heritage) by microwave-assisted process (MAP) was studied. A central composite rotate design (CCRD) was used to obtain the optimal conditions of microwave-assisted extraction (MAE), and the effects of operating conditions such as the ratio of solvents to materials, microwave power and extraction time on the extraction yield of Acys were studied through response surface methodology (RSM). The optimized conditions of MAE were ratio of solvents to materials 4:1 (ml/g), extraction time 12 min, and microwave power 366 W. Under these conditions 43.42 mg of Acys from 100 g of fresh fruits (T _Acy, expressed as cyanidin-3-glucoside), approximately 98.33% of the total red pigments, could be obtained by MAE. The Acys compositions of extracts were identified by high-performance liquid chromatography – mass spectrometry (HPLC-MS), 12 kinds of Acys had been detected and 8 kinds of Acys were characterized. Result indicated that cyanidin-3-sophoroside, cyanidin-3-(2 ^G -glucosylrutinoside), cyanidin-3-sambubioside, cyanidin-3-rutinoside, cyanidin-3-xylosylrutinoside, cyanidin-3-(2 ^G -glucosylrutinoside), and cyanidin-3-rutinoside were main components in extracts. In addition, in comparison with the conventional solvent extraction, MAE is more efficient and rapid to extract Acys from red raspberry, due to the strong disruption of fruit tissue structure under microwave irradiation, which had been observed with the scanning electron microscopepy (SEM). However, the Acys compositions in extracts by both the methods were similar, which were investigated using HPLC profile.     

利用中压制备液相色谱从桑葚中快速制备矢车菊素-3-葡萄糖苷单体

单体花色苷的快速大量制备长久以来是花色苷产业化中的难题,而中压制备液相色谱在产业应用中有着很大的开发空间。选取花色苷组分较单一的桑葚为实验原料,经提取分离总花色苷后使用填装有反相C18填料的耐压玻璃柱作为中压制备液相色谱柱,纯化制备矢车菊素-3-葡萄糖苷单体。结果显示：3 个色谱分离峰中目的峰（峰2）经高效液相色谱和质谱确证为由矢车菊素-3-葡萄糖苷（cyanidin-3-glucoside,C3G）和矢车菊素-3-芸香糖苷（cyanidin-3-rutinoside,C3R）组成,采用峰面积归一化法计算得到C3G纯度为73.56%;通过对峰2采用切割方式进行收集,C3G纯度达到98%以上,单次收集到C3G单体溶液650?mL。中压制备液相色谱法单次上样量大、步骤简洁、成本低廉,可为矢车菊素-3-葡萄糖苷单体的规模化生产提供一定的参考。     

Identification of phenolic compounds isolated from pigmented rices and their aldose reductase inhibitory activities

Two anthocyanins (cyanidin-3-O-β-glucoside and peonidin-3-O-β-glucoside) and other phenolic (ferulic acid) were, respectively isolated from black and pigmented brown rices (Oryza sativa L. japonica) and their complete structures were determined by spectroscopic analyses (H NMR, C NMR and MALDI MASS). The HPLC profile of anthocyanins extracted from black rice showed cyanidin-3-O-β-glucoside as the first peak (85%) and peonidin 3-O-β-d-glucoside as the second (15%), while that of pigmented brown rice showed ferulic acid as the first peak (85.7%) and tocols as the second (14.3%). Several tocols were isolated and identified from the unsaponifiable fractions of both rices having some difference on their structures and amounts. The aldose reductase inhibitory activity of isolated compounds was in the following decreasing order: cyanidin-3-glucoside > quercetin > ferulic acid > peonidin-3-glucoside > tocopherol.     

The chemistry and medicinal uses of the underutilized Indian fruit tree Garcinia indica Choisy (kokum): A review

Garcinia indica Choisy Syn Brindonia indica, commonly known as kokum and belonging to Guttiferae family, is a plant native to certain regions of India. The trees yield fruits annually in the summer season during the months of March to May. The fruits are green when raw and red to dark purple when fully ripe. They are used to prepare juice, pickles and as acidulant in curries. In the traditional Indian system of medicine the Ayurveda and in various folk systems of medicine, the fruit rinds and leaves are used to treat various inflammatory ailments, rheumatic pain and bowel complaints. The kokum butter prepared from the seed is of both commercial and medicinal use. Chemical studies have shown that the rind contains protein, tannin, pectin, sugars, fat, organic acids like (−)-hydroxycitric acid, hydroxycitric acid lactone and citric acid; the anthocyanins, cyanidin-3-glucoside and cyanidin-3-sambubioside; and the polyisoprenylated phenolics garcinol and isogarcinol. Preclinical studies have shown that kokum or and some of its phytochemicals possess antibacterial, antifungal, anti-ulcerogenic, cardioprotective, anticancer, chemopreventive, free radical scavenging, antioxidant and anti-obesity effects. The present paper reviews the nutritional value, the phytochemical compounds, traditional uses and validated pharmacological properties of kokum.     

Effects of pectolytic enzyme treatments on anthocyanins in raspberry juice

Raspberry juices treated with two commercial pectinase preparations lost almost 20% of their total anthocyanin pigments; losses were related to both concentration of enzyme and time of treatment. Analysis of individual anthocyanins showed differences between the enzymes. Ultrazyme reduced all components in similar proportions, whereas Pectinex apparently also converted cyanidin-3-sophoroside and cyanidin-3-glucosylrutinoside into cyanidin-3-glucoside and cyanidin-3-rutinoside respectively, presumably by hydrolysis of β1–2 glucosidic bonds by glucosidase present in the Pectinex.     

Chemical composition and antioxidant activity of tronchuda cabbage internal leaves

A phytochemical study was undertaken on the internal leaves of tronchuda cabbage (Brassica oleracea L. var. costata DC). Seventeen phenolic compounds were characterized and quantified by reversed-phase HPLC-DAD-ESI-MS ⁿ and HPLC/DAD, respectively: quercetin 3-O-sophoroside-7-O-glucoside, 3-p-coumaroylquinic acid, kaempferol 3-O-sophoroside-7-O-glucoside, kaempferol 3-O-(caffeoyl)-sophoroside-7-O-glucoside, sinapoyl gluc-oside acid, kaempferol 3-O-(sinapoyl)-sophoroside-7-O-glucoside, kaempferol 3-O-(feruloyl)-sophoroside-7-O-glucoside, kaempferol 3-O-(p-coumaroyl)-sophoroside-7-O-glucoside, 4-p-coumaroylquinic acid, sinapic acid, kaempferol 3-O-sophoroside, 3 isomeric forms of 1,2-disinapoylgentiobiose, 1-sinapoyl-2-feruloylgentiobiose, 1,2,2-trisinapoylgentiobiose and 1,2′-disinapoyl-2-ferul-oylgentiobiose. Seven organic acids (aconitic, citric, ascorbic, malic, quinic, shikimic and fumaric acids) were also identified and quantified. The hot water extract of tronchuda cabbage internal leaves was investigated for its capacity to act as a scavenger of DPPH^• radical and reactive oxygen species (superoxide radical, hydroxyl radical and hypochlorous acid), exhibiting antioxidant capacity in a concentration dependent manner against all radicals.     

Mechanisms of anthocyanin degradation in grape must-like model solutions

Oxidative degradation of o-diphenolic (eg cyanidin-3-glucoside) and non-o-diphenolic (eg malvidin-3-glucoside) anthocyanins in the presence of caffeoyltartaric acid and grape polyphenoloxidase was studied in model solutions. Both anthocyanins reacted with the enzymically generated caffeoyltartaric acid o-quinone. Kinetic studies indicated that cyanidin-3-glucoside was degraded mostly by coupled oxidation whereas malvidin-3-glucoside formed adducts with caffeoyltartaric acid quinone. In solutions containing equimolar amounts of the anthocyanins, both reactions took place but coupled oxidation of cyanidin-3-glucoside occurred at a faster rate, partly protecting malvidin-3-glucoside. The occurrence of coupled oxidation resulted in sparing of caffeoyltartaric acid. Several oxidation products, including red pigments, were detected by HPLC. The UV-Vis spectra of the coloured condensation products suggested that they contained both caffeoyltartaric acid and anthocyanin moieties. All these pigments were gradually degraded to colourless compounds either by enzymatic oxidation or by further reaction with quinones.     

Flavonoids, sugars and fruit acids of alpine bearberry (Arctostaphylos alpina) from Finnish Lapland

Alpine bearberry (Arctostaphylos alpina L.) is a special circumpolar edible berry primarily used for household purposes but with greater potential in commercial applications than utilized thus far. In this study, the flavonoids of alpine bearberry were investigated with HPLC-DAD and HPLC-MS, and the sugars and fruit acids as trimethylsilyl derivates with GC-FID. The most abundant anthocyanin was cyanidin-3-O-galactoside accounting for over 95% of the anthocyanins. Eight other anthocyanins existed in trace amounts only, the richest of them being cyanidin-3-O-arabinoside and cyanidin-3-O-glucoside. Cyanidin accounted for over 99% of the anthocyanin aglycones. All the flavonols identified were glycosides of quercetin. Majority of the sugars were glucose and fructose while the most abundant fruit acid was quinic acid. The unique anthocyanin composition of alpine bearberry encourages advanced use of the berry as a food ingredient in private households and in industrial applications. In addition, it serves as a unique reference material for research purposes.     

Effect of cyclodextrins on the thermodynamic and kinetic properties of cyanidin-3-O-glucoside

In this study, the effect of α and β-cyclodextrin on cyanidin-3-O-glucoside color was investigated by UV–visible absorption techniques. The equilibrium and kinetic constants of the network of chemical reactions taking place in cyanidin-3-O-glucoside were also studied in water at 25 °C by UV–visible absorption techniques. The results showed that the addition of β-cyclodextrin resulted in the fading of anthocyanin solution, and this fading effect was greater at higher pH. This anti-copigmentation effect is caused by the selective inclusion and stabilization of the anthocyanin colorless forms into the β-cyclodextrin cavity. Oppositely, no changes were observed in the cyanidin-3-O-glucoside absorption spectra with the addition of α-cyclodextrin.Direct pH jump, from thermally equilibrated solutions at pH = 1.0 (flavylium cation, AH⁺), shows three kinetic processes: formation of the base A, hydration reaction to form the hemiketal B and the chalcone cis–trans isomerization (Cc–Ct). The results obtained clearly indicated that the equilibrium and kinetic constants of the network of chemical reactions taking place in cyanidin-3-O-glucoside were affected by the presence of β-cyclodextrin. Molecular inclusion in the β-cyclodextrin cavity resulted in the increase of the isomerization observed rate constant (k_obs) at pH 5.3 and in the increase of the hydration equilibrium constant K_h which is in agreement with the fading of the anthocyanin solution. For the macrocycle α-cyclodextrin, no significant changes were observed on the equilibrium and kinetic constants, which suggests that the inclusion of cyanidin-3-glucoside in the α-cyclodextrin's cavity is not favored.     

Chemistry and Quality of Hibiscus (Hibiscus sabdariffa) for Developing the Natural-Product Industry in Senegal

ABSTRACT:  The objectives of this study were to assess and improve the quality of the hibiscus calyces from Senegal over 2 production seasons (2004 to 2005), to develop and adapt new procedures for the determination of hibiscus anthocyanins and analysis of the 2 major ones, delphinidin-3-sambubioside and cyanidin-3-sambubioside. The foreign matter, total ashes, and acid insoluble ashes showed that the calyces harvested in 2005 were produced following hygienic practices, while the color assessment of the calyces and analysis of hibiscus active principles also showed higher amounts of anthocyanins in 2005. A protocol to measure anthocyanins by pH-differential UV-Vis spectrophotometry was adapted to measure the hibiscus anthocyanins from a water extract. The spectrophotometric method for quantitation of total anthocyanins showed a close correlation ( r ²= 0.82) when compared with the HPLC method, suggesting the use of the colorimetric method in quality control programs as an affordable alternative method to assess anthocyanin content in hibiscus. New and raised standards for the cleanliness and active principle content in hibiscus are also proposed. This study demonstrated that the implementation of a quality control program and the application of agricultural good practices in the production and processing of hibiscus calyces can lead to higher quality natural plant products.     

Characterisation of anthocyanins from Garcinia indica Choisy

The present study deals with isolation and characterisation of anthocyanins present in Garcinia indica Choisy (popularly known as kokum), which is a potential source of a natural food colourant. The kokum was found to contain a very high concentration of anthocyanins (2.4 g/100 g of kokum fruit), compared to other natural sources. Acid hydrolysis ascertained that this anthocyanin consisted of a single aglycone, i.e., cyanidin. A comparison of saponified and unsaponified fraction indicated that the pigment was not acylated. HPLC, mass and NMR spectroscopy analyses confirmed that the pigment essentially contains two anthocyanins, which were identified as cyanidin 3-glucoside and cyanidin 3-sambubioside.     

First accurate profiling of antioxidant anthocyanins and flavonols of Tibouchina urvilleana and Tibouchina mollis edible flowers aided by fractionation with Amberlite XAD-7

A purification and fractionation process of the edible flowers of Tibouchina mollis and Tibouchina urvilleana followed by the first attempt to the anthocyanin and flavonol characterisation and identification by UHPLC-DAD-ESI-MS were developed. T. urvilleana exhibited a higher monomeric anthocyanin content, mainly due to the presence of the 3-O-(6′-p-coumaroyl)-glucoside derivatives of malvidin and petunidin. Quercetin-3-O-hexoside was the major flavonol identified in T. urvilleana, and the lack of myricetin derivatives was also exhibited. The anthocyanin and flavonol profile of T. mollis was more miscellaneous, characterised by the occurrence of cyanidin-3-O-glucoside followed by the 3-O-(6′-p-coumaroyl)-glucoside and 3-O-glucoside derivatives of malvidin and petunidin as anthocyanins, and myricetin, quercetin, and 3-O-hexosides of kaemperol and quercetin as flavonol compounds. Therefore, the anthocyanin and flavonol profile, through a process based on purification and fractionation, could be a useful tool to ensure the authenticity of the Tibouchina. Furthermore, the purification process made the antioxidant activity increase, which is greatly correlated to the reduction capacity.